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Cryo-mesh: a simple alternative cryopreservation protocol
B Funnekotter1, E Bunn2, R L Mancera3
1School of Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, Perth; Botanic Gardens and Parks Authority, West Perth, Australia.
Background:
The continued development of new cryopreservation protocols has improved post-cryogenic success rates for a wide variety of plant species. Methods like the cryo-plate have proven beneficial in simplifying the cryopreservation procedure.
Objective:
This study assessed the practicality of a stainless steel mesh strip (cryo-mesh) for cryopreserving shoot tips from Anigozanthos viridis.
Materials And Methods:
Shoot tips of A. viridis (Kangaroo Paw) were precultured on 0.4 M sucrose medium for 48 h. Precultured shoot tips were coated in a 2% alginate solution and placed onto the cryo-mesh (a 25 x 7 mm, 0.4 mm aperture, 0.224 mm diameter wire stainless steel mesh strip). The alginate was set for 20 min in a loading solution containing 100 mM CaCl2, anchoring the shoot tips to the cryo-mesh. The cryo-mesh was then transferred to PVS2 on ice for 20, 30 or 40 min prior to plunging the cryo-mesh into liquid nitrogen. The cryo-mesh protocol was compared to the droplet-vitrification protocol.
Results:
A maximum of 83% post-cryogenic regeneration was achieved with the cryo-mesh when exposed to PVS2 for 30 min. No significant difference in post-cryogenic regeneration was observed between the cryo-mesh and droplet-vitrification protocols.
Conclusion:
Anigozanthos viridis shoot tips were successfully cryopreserved utilising the new cryo-mesh. The cryo-mesh thus provides a simple and successful alternative for cryopreservation.
